CZ270094A3 - Method of determining an activity stage of stone formation and composition of stone-forming salts of urea at urolithiasis - Google Patents

Abstract

The method for determining the degree of lithogenesis activity and the composition of lithogenic urine salts comprises adding appropriately to a sample of urine an aqueous protein solution in a ratio of 9:1, 7:1 or 5:1, applying the resulting mixture in the form of a drop to a flat surface and drying within at least 24 hours, then determining the mode of salt crystallisation in the edge zone of the sample, concluding whether the degree of lithogenesis activity is weak, moderate or pronounced in the presence of single crystals, conglomerates thereof or complete crystallisation of this zone, and determining the composition of these crystals formed with parallel determination of the salt composition in the central zone of the sample and on later performance of a comparative analysis of these compositions for the purpose of determining the lithogenic urine salts.

Description

Method for determining the degree of stone formation activity and composition of urinary stone-forming salts in urolithiasis

Technical field

The method of determining the stone formation activity and the composition of the urinary stone-forming urine in urolithiasis.

BACKGROUND OF THE INVENTION

At present, problems of dynamic research process calculus formation in the human organism, its intensity, determining the composition of salts in the process of the stone appears to be _the current as possible to prevent urinary stone formation, while said control process and to determine and implement individual therapy in urolithiasis.

Methods for predicting urolithiasis based on the discovery of a stone formation process are known, for example, a method of prognosticization based on the appearance of crystals in fresh urine or soon after urination / ¥. Pre-treatment: Clinical laboratory follow-up 19 ^ 4, Medicine (pp. 420-446), or a method for predicting urolithiasis (AO SU 1629846), based on the determination of rat urinary formation activity by mixing solutions of tannic chloride, sodium oxalate and urine, exposure and counting the crystals formed.

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T T <y A X - - «i Λ ** ** t»., «. uveucuc ajjuouuj jouu stones and do not allow to assess the degree of activity of this process.

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A method for predicting urolithiasis is known in which it is proposed to determine the degree of risk of crystallization of calcium phosphate in urine by calculating formed crystals of a certain size by mathematical calculation (Urological Research No. 2,15, 1987, Springe Verlag; H.-G, Tiselius; Measurement of Risk of Calcium Phosphate Crystallization in Urine, pp. 79-81 /. This method is complicated by mathematical calculations and is also aimed only at detecting the stone formation process.

A method for prognosticating urolithiasis (PCT SU 91/00140) is also known in which a protein solution, such as albumin, is added to the urine sample, the mixture is dried and urolithiasis is prognostic when the sample is crystallized. Said method is only aimed at detecting the stone formation process and cannot be used to determine the degree of stone formation activity. *

In addition, methods for determining the salt composition of urinary thiase (V.E.) are known. Rukovodstvo po kliničeskim laboratornom issledovanijam 1964 Medicina Moskva, pp. 452–454; Y / andt, M, Underhill, L .: Brit. J., Urol. 61, 1988, No. 6, pp. 478-481;

Schubert ,. G., Brien, G., Adam, K .: Z. Clin., Med. 44. 1988, No. 11, pp. 923-928; Nichino, T., Sakura, T., Sato, T., Koiso, K., Kneknek, S .; "Me p. J. Neptirol., 1987, No. 5, pp. 571-575].

All these methods are based on urine detection .........

(deposition of salts, sand, stones) by a physicochemical route and the use of adsorption, ion exchange, for example chromatography, X-ray spectral analysis, magnetic analysis using magnetonuclear resonance, electron paramagnetic resonance or other spin effects.

These methods are suitable only for detecting the formation of stones excluded from the organism and do not allow to determine the chemical character of the formation of stones in the early stages of urolithiasis, or formed but not excluded from the organism.

The method of determining the chemical composition of stone-forming salts in the early stages of urolithiasis and the resulting, but not secreted stones, is unknown.

It provides the invention

The method of determining the degree of stone formation activity and the composition of urinary stone-forming salts in urolithiasis according to the invention appears new and is not described in the literature.

The object of the invention is to provide such a method which would be useful. - to determine rapidly and accurately the degree of stone formation activity in urolithiasis as well as to determine the composition of stone-forming salts at any stage of their formation.

According to the invention, an aqueous protein solution in the ratio of 9: 1, 7: 1 or 5: 1 is added to the urine sample according to the invention in the method for determining the stone formation activity and composition of urinary stone-forming salts in urolithiasis. The droplets are applied on a flat surface and dried for at least 24 hours, whereupon the characterization of the salt crystallization in the edge zone is determined. The presence of single crystals, conglomerates thereof or complete crystallization in this zone is judged to be a weak, adequate or significant degree of stone formation activity and the composition of these crystalline bodies is determined while determining the composition of the urine-forming urine salts. * ·. . ·

-and*

As an aqueous protein solution, preferably an 8-12% aqueous albumin solution is used. In order to 'increase the accuracy' of the analysis, the drying of the droplet of the mixture applied on a flat surface is expediently carried out at ambient temperature. To accelerate and increase the accuracy of the analysis, to determine the elemental composition of the crystalline formations; preferably X-ray spectral microanalysis is used. The method according to the invention revealed the dependence of the stone formation activity and the intensity of salt crystallization in the marginal zone of the urine sample. .

This allows to quickly and accurately determine the level in the urine sample of stone formation activity, allowing Drovádět R * stone formation process from the standpoint of dynamics, simultaneously fed to said pro- _su correction, thus preventing crystallization of urinary stone, and as appropriate, to implement individual therapy . In addition, the method according to the invention allows, before the formation of stone in the body, to determine the composition of the rock-forming salts and to take remedial measures to eliminate them from the body and to prevent the progress of these salts with drinking water and nutritional products. The method of analysis according to the invention allows the discovery of stones in the patient's urinary tract, over a period of observation, to identify which stone-forming salts form their surface layer, which may be necessary, for example, in lithotrypsia.

Monitoring the composition dynamics of the rock-forming salts (including from the beginning to their formation) provides the possibility to determine the composition and the subsequent structure of the whole stone, which is important. the implementation of the "mode, if -________ ₂ totrypsia."

Moreover, the method of analysis according to the invention makes it possible to realize effective prophylaxis of recurrence of urolithiasis. The method is simple, simple to implement, requiring no special instrumentation.

The method of determining the degree of stone formation activity of the present invention is based on the observed dependence of the degree of salt crystallization on the edge layer of the urine sample on the stone formation activity. The process according to the invention is carried out as follows;

An aqueous protein solution is added to the urine sample (preferably 8-12% aqueous albumin solution is used as such) in a ratio of 9: 1, 7: 1 or 5: 1. A urine sample with an aqueous protein solution may be used for analysis. any of the above ratios. ^{"5A; toi} ^ o '^ 8pólO'-hllyó ≤ t ≤ anal-ZYM .-« ^ © € the account of «^ dv neboutěi- acianty ^ ^ y ^ s ^ out by ratios. The beaker is then applied to a flat surface and dried for at least 24 hours at ambient temperature.

After drying, the intensity of crystallization of the urine salts in the edge layer of the sample is determined: The individual crystals indicate a weak activity of the stone formation process.} The occurrence of conglomerates of salt crystals indicates an adequate activity; complete crystallization of the salts of this zone shows a marked activity of the stone formation process.

The absence of crystals in the peripheral zone indicates that the formation of stones does not occur. The method according to the invention makes it possible simultaneously to determine the degree of activity i of the stone-forming salts of the mash. When crystalline formations occur in the peripheral zone of the sample, by means of. According to the methods (preferably X-ray spectral analysis), the elemental composition of these crystalline structures is determined. At the same time, the composition of the salts of the central portion of the urine sample is determined and a comparative analysis of these compositions is made, which concludes on the chemical composition of the stone-forming salts. Stone formation is a dynamic process whose salt content can change over time. The method according to the invention makes it possible to determine the content of individual salts in the cation which arises, or can be formed prospectively, at a given urinary salt content.

The method of the invention has been tested in clinical laboratories. To determine the degree of stone formation, 12 patients were observed in which Deriodic was observed to spontaneously disrupt and eliminate stones, and 128 practically healthy subjects in the subclinical stage of urolithiasis, as determined by a previously known method, but who did not know the degree of stone formation. Research consisted of the following: Urine samples were taken, to which an aqueous solution of protein (albumin) was added at a ratio of 9: 1, 7: 1 and 5: 1, respectively. a drop of the obtained mixture was deposited on the slide surface and dried for 24 hours. The presence of three crystals was then detected in the peripheral zone of the dried sample. The research results are shown in Tables 1 and 2,

As can be seen from the results shown in Tables 1 and 2, 100% agreement of laboratory data with clinical stone formation was found. Stone formation was observed in 2-3 months in one group of patients who experienced stone excretion with a significant degree of stone formation activity (complete marginal crystallization); in the second group, with partial crystallization of the edge zone / crystal conglomerates /

- adequate degree of stone formation activity - stone formation was found in 7-8 months ;, in the third group with single crystals in the marginal zone (weak degree of stone formation activity) stone formation was found in 2-2.5 years. The same can be said for clinical and laboratory data in practically healthy subjects in the subclinical stage of urolithiasis (Table 2). Thus, the method of the invention allows to conclude on the intensity of the Stone formation process (to determine the degree of activity) as well as the times of stone formation.

Simultaneously with the determination of the degree of stone formation activity in 13 patients with urolithiasis (with a marked degree of stone formation activity), the composition of stone-forming salts was monitored. Of these 13 patients, 6 were self-excreting stones, 7 patients were subjected to lithotripsy. From a specimen dried on a glass slide, which showed complete marginal crystallization (indicative of a significant degree of stone formation), was performed by X-ray structure Table 1

Clinical and laboratory data of patients secreting stones

¹ 1, 1 Item no. I '1 i i Investigation Urine / aqueous solution of aluminium Level of activity: creation , stones F Stone discovery times in subsequent clinical investigations / months / 9: 1 7: 1 It was 5 - 1 at the break 1 2 3 4 5 6 7 1 Patient B. + 1 1 1 weak 27 Mar: 2 Patient L. + - - weak 24 J Patient Ja. « - weak _ _ ______29 ---- 4_ ýpac ien-t ^ Š . ^ +++ - - --- fcfc-  - -direct- 5 Pacient M. ++ - appropriate 8 6 Pacient R. +++ ++ - appropriate ⁸ 7 Patient 0 .. +++ ++ - appropriate 7 8 Pacient K. +++ ++ - appropriate 9 9 Patient S. +++ ++ - appropriate 8 10 Patient U. +++ ++ • - adequately 7 11 Patient A. +++ +++ +++ distinctive 3 12 Patient D. +++ +++ +++ distinctive 2

Note: + - presence of individual crystals in the edge zone of the sample ++ - crystal conglomerates +++ - total crystallization of the edge zone in

’I.

- Ί Table 2.

Clinical and laboratory data in practically healthy subjects in the subclinical stage of urolithiasis

Ref. Number examinations- vaných persons Urine / aqueous albumin solution ratio Degree activities creation stones Periods of stone finding in subsequent clinical examinations / months / 9: 1 7: 1 It was 5 - 1 at the break 1 2 3 4 5 6 7 1 61 + weak 24-30 2 49 +++ ++ - appropriate 7-8 3 18 +++ +++ +++ distinctive 2-3

Note: + - presence of individual crystals in the edge zone of the sample ++ - crystal conglomerates +++ - total crystallization of the edge zone

Table 3

Comparative data of phase composition analysis of crystalline marginal Diasma specimens of urine sample and precipitated stones

, No. Investigations and- and and Diagnosis Composition / elemen- Composition of excretory, phase / urinary urine salts of crystalline marginal specimens of the urine sample currencies / prevailing • salt in surface ΐ ‘ tier / 1 2 3 4 5 : 1 Patient TO. Uric acid ⁺ Udrs 2 Patient P. Uric acid, ammonium urate ⁴ “ Udrs i 3 Patient Ch. Uric acid ₊ acid, ammonium urate Udrs i ⁴ ' Patient -L. - Uric acid- Udrs í 5 Patient R. Secondary Calcium, phosphorus, magnesium ⁺⁺ Phosphates ! nyelonefrit - 6 Patient AND, Secondary Dyelonefrit Vánhík ,, phosphorus, magnesium Phosphates :? and and Patient ř _t .Secondary Dyelonefrit Calcium, phosphorus, zinc Phosphates j 8 j ' ΐ Patient IN. Secondary Dyelonefrit ' Phosphorus ⁺⁺ Phosphates ! 9 Patient AND. .... _ Phosphorus .... calcium, magnesium '** Phosphates. . ko Patient AND. . Phosphorus, Calcium ** Phosphates ; 11 Patient G. Secondary Calcium** Oxalates i ‘ pyelonephrite 'I ² Patient T. Secondary Calcium Oxalates 4 - ------. - Dyelonefrit ............... <* i 13 . Patient A. Secondary Calcium Oxalates oyelonefrite and

Note +. - X-ray structure analysis data ++ - X-ray spectral analysis data

- 9 tour analysis, X-ray spectral microanalyse determination of phase and elemental composition of these crystalline formations.

The obtained data of the elemental (phase) composition of the crystalline marginal specimens of the samples was compared with the data of the analysis of the subsequently excreted urinary stones. Monitoring results are presented in Table 3.

As can be seen from the results presented in Table 3, the data ^- “* · * - ¹ '· - 41. i ·. _B * j *. ·, The analysis of the crystalline marginal structure of the urine sample corresponds to the data obtained from the urinary calculus *.

In this way, the method of the invention makes it possible to predict or determine the phase composition of the stone in the subclinical stages of urolithiasis development, which can be used to prevent the formation of stones.

For a better understanding of the present invention, the following examples of specific variations of the method of the invention are given.

DETAILED DESCRIPTION OF THE INVENTION

Example 1.

Patient B., 50, was supervised by a clinical laboratory for 2 years. It was taken under dynamic observation after the stone was removed from the body. Weekly urine monitoring was performed as described. The monitoring methodology consisted of adding an aqueous solution of protein at a ratio of 9: 1, T: 1 or 5: 1 to the urine sample. The droplets of the obtained mixture were deposited on a glass slide and dried at ambient temperature for 24 hours. Thereafter, the crystallization pattern of the salts in the sample zone of the sample was determined. Within four months there was a significant degree of stone formation activity (complete crystallization of the sample edge zone). Simultaneously with the aid of spectral microanalysis, the elemental composition of crystalline boundary band formation was found. For the purpose of comparative analysis, the determination of the elemental composition of the center band of the sample was simultaneously performed. A considerable increase in the content of the individual elements in the marginal zone with respect to the central zone forms the basis of the stone-forming salts.

The results of the determination of the composition of stone-forming salts were confirmed by analysis of the composition of the stone after its excretion from the organism.

Monitoring results are presented in Table 4.

Table 4

Comparative analysis data of the elemental composition of the central and peripheral zones of the urine sample and the excreted stone

Ref. Material P r v k y in % On Mg P WITH Cl TO Ca Zn Si 1 2 3 4 5 6 7 8 Urine Sample / Middle. zone/ 27.4 . i, i 3.4 1.1 62.6 3.3 - 2 Urine / marginal sample zone/ 17.3 0.9 14.6 9.5 39.6 11.6 56,1 - - Excreted urinary stone - 3.3 30.3 - 4 » - 64,2 - -

As can be seen from the data in Table 4, the stone-forming elements appear Ca and P, which is confirmed by the composition of the stone (after its exclusion from the body).

After removal of the stone from the body, the patient was given medical treatment and prolonged weekly monitoring of urine in this way, which was performed for two years. Dynamic observation / periodic determination of the degree of stone formation activity / patient is presented in Table 5 ·

Table 5

Results of monitoring the dynamics of the stone formation process in patient B. after treatment initiation

Tracking period Characterization of the marginal zone of urine sample at urine / aqueous solution ratio albumin Degree activities creation stones 9: 1 7: 1 It was 5 - 1 at the break 1 2 3 4 5 1 week ++ ++ - appropriate 1 month + + * · » weak 2-4 months, in. -. does not take place 5-7 months ++ + - appropriate 8 months + - - weak 9-13 months - - - does not take place 14 months + - - weak '15 -20 months - - weak

Note: + occurrence of single crystals ++ crystal conglomerates

- without crystals

Example 2.

Determining the degree of stone formation activity in a patient's urine sample Ch. was performed analogously to the procedure described in Example 1. A significant degree of stone formation activity was observed (complete crystallization of the sample edge zone). Elemental composition of crystalline boundary band formation was determined by X-ray spectral microanalysis. A comparative analysis of the elemental composition of the marginal and central zones of the dried urine sample was performed. The results of the determination of the composition of the stone-forming salts were confirmed by the analysis of the composition of the stone after its elimination from the organism. The monitoring results are presented in Table 6.

Table 6.

Comparative analysis data of the elemental composition of the central and peripheral zones of the urine sample and of the excluded stone

No. Material Elements v % On Mg P S Cl TO Ca Zn Si 1 2 3 4 5 6 7 8 9 10 11

1 * Sample-urine central 26.2 band

2. Marginal urine sample 27.6

--- band ------ - ~

Excreted urinary stone, stone * ⁷

2,0 15,1

2, JL 19.6

8.9 30.5

12,3 26,0

13.0

4.8

4.1

0.5 2.6 7.6 - 93.0 -. As can be seen from the data of Table 6, the main element of the stone-forming salts appears to be mainly Ca, which was confirmed by the composition of the stone: Ca = 93% ·

Example 3.

The determination of the degree of stone formation activity in the urine sample of patient K. was performed analogously to the example Ί. A significant degree of stone formation activity (complete marginal crystallizer of the specimen) was found. Elemental rat composition was determined by X-ray spectral microanalysis. of the marginal zone. A comparative analysis of the elemental composition of the marginal and central zones of the dried urine sample was performed. The results of the determination of the composition of the stone-forming salts were confirmed by the composition of the stone after its exclusion from the organism. The monitoring results presented in Table 7 ·

Table ?.

Comparative analysis data of the elemental composition of the central and peripheral zones of the urine sample and the excreted stone

"" 4 .__; ; Elements in% L · X O · Jun in Cl x 10x On Mg P S Cl TO Ca Zn S · * ** - Λ , - -Γ 1. Central urine sample. . zone 17.2 - 15,8 12,2 31.1. 19.7 2,6 - - 2. Urine sample edge zone 12.7 . 3.9 23,3 9,2 22.4 12,5 .. , .5,5. - - 3 * Excreted urinary stone - 14.0 39.3 - - 44.7 - - -J *

As can be seen from the data in Table 7, the elements of the stone-forming salts appear essentially to be Ca, P and Mg, which is confirmed by the composition of the stone: Ca - 44.7 P = 39.3% and Mg = 14.0

Example 4.

Determination of the degree of stone formation activity in the urine sample of patient S *. A significant degree of stone formation activity (complete crystallization of the marginal zone) was found. By means of X-ray spectral microanalysis, the elemental composition of the crystalline boundary band structures could be determined. A comparative analysis of the elemental composition of the boundary and midband of the dried urine sample was performed. After finding a significant degree of stone formation activity and determining the composition of stone-forming urine salts, the patient received a course of individual therapy. A secondary analysis of the urine sample was performed. The monitoring results are presented in Table 8.

Table 8.

Comparative analysis data of the elemental composition of the middle and peripheral zones of the urine sample and the precipitated stone

Monitoring period

Degree of stone formation activity

Material

Elements

Na Mg P S Cl *

To Ca Zn Si

3. 4 5 6 7 8 9 10 11 12

Significant before treatment

Central urine sample

25.2 1.5 7.1 7.8 37.1 16.4 1.7 0.2 Urine sample marginal band

14.1 0.214.6 2.7 17.0 8.4 4.7 0.2 After treatment

Sample

The urine process does not run through the central zone

Urine sample marginal band

33.4. 1,0 10,6 10,, 1 35,9 7,1 1,4 1,3 34,6 - 10,6 8,3 35,9 6,4 0,4 1,, 1 As can be seen from the table 8, Ca and F appear to be the main elements of the stone-forming salts (prior to ¹ treatment); after treatment stone formation process does not take place - - - Industrial applicability

The method of the invention can be used in clinical and laboratory studies to analyze the degree of stone formation activity to follow the dynamics of stone formation processes, to determine the phase composition of the stone in the early stages of urolithiasis to prevent urinary stone formation and to determine individual urolithiasis therapy.

Claims (4)

PATENT CLAIM! A method for determining the degree of stone formation activity and the composition of urinary stone-forming salts in urolithiasis, characterized in that an aqueous protein solution in a ratio of 9: 1 is added to the urine sample. 7: 1 or 5: 1, the mixture obtained is applied in a dropwise manner to a flat surface and dried for at least 24 hours. whereupon the character of the crystallization of the salts in the marginal zone of the sample is determined and, in the presence of the individual crystals, their conglomerates or the complete crystallization of this zone, a weak _T 'adequate or significant degree of stone formation activity is judged; the composition of these crystalline formulations is determined with simultaneous determination of the salt composition of the central zone of the sample, and a comparative analysis of these compositions is then performed to determine the stone-forming urine salts. Method according to claim 1, characterized in that an aqueous to albumin solution of between 12 and 12% is used as the aqueous protein solution. 3. The method according to one sheet, characterized in that drying of droplets získanér.émšsi, _of: applying on a flat surface, is performed at ambient temperature. Method according to one of claims 1 to 3, characterized in that X-ray spectral microanalysis is used to determine the elemental composition of the crystalline formations.